DSMZ 193a . PHB/PYRUVATE MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
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Selected strain: unclassified bacterium DSM 6754

This strain uses the standard medium without modifications.

Equipment needed: Autoclave Gassing station Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 942 ml - -
Solution B 30 ml - -
Solution C 20 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
1 Solution A is sparged with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere in anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution E is autoclaved separately under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 6.8 - 7.0.
2 After inoculation, pressurize vessels with 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.185 22.421
KH2PO4 0.20 g 0.212 1.56
NH4Cl 0.30 g 0.318 5.954
NaCl 7.00 g 7.431 127.156
MgCl2 x 6 H2O 1.30 g 1.38 6.788
KCl 0.50 g 0.531 7.12
CaCl2 x 2 H2O 0.15 g 0.159 1.083
Selenite-tungstate solution 1.00 ml - -
Trace element solution SL-10 1.00 ml - -
Poly[(R)-3-hydroxybutyric acid]
(SIGMA)
1.00 g 1.062 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.3e-4 0.002
Distilled water 940.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate 5 g 250 2271.82
Distilled water 20 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.4 g 40 166.54
Distilled water 10.0 ml - -
Solution occurs in 176 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
Solution occurs in 263 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%)
10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
Solution occurs in 299 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % H2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 1
unclassified bacterium DSM 6754

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 6.979 119.423
Na-pyruvate 4.985 45.301
Na2SO4 2.991 21.058
Na2CO3 1.496 14.11
MgCl2 x 6 H2O 1.296 6.375
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Cultivation metadata from 1 strains BacDive

oxygen anaerobe 1 strain
temperature mesophilic 1 strain